News Resource: School of Life Science

Editor: News Agency of BIT

Translator: Bo Yilin, News Agency of BIT

Recently, Professor Huo Yixin’s team in the School of Life Science has made important research results on the modification and application of advanced alcohol biosensors, which was published in ACS Synthetic Biology, a top journal in the first area of Biochemical Research Methods category. The first authors of this paper are PhD student Wu Tong and associate researcher Chen Zhenya, and the corresponding author is Prof. Huo YiXin.

Advanced alcohols such as n-butanol, isobutanol or isoamyl alcohol are a promising class of bioenergy alternatives to fossil fuels due to their high energy density and low hygroscopicity, as well as their low corrosiveness and non-volatility. In recent years, a lot of work has been done to dynamically monitor the yield of advanced alcohols through labeling and screening systems under continuous sampling conditions, but these methods still cannot achieve real-time monitoring and are easy to contaminate samples; In contrast, biosensing systems have higher safety, stability and universality, and are more suitable for continuous monitoring in industrial production processes.

Biosensors have been widely used in the screening of high-yielding strains for metabolic engineering and in the regulation of metabolic pathways. By designing, constructing and modifying biosensing systems to dynamically respond to changes in signal substance concentrations and initiate cellular output, thus, biosensors are a tool for rapid, high-throughput assessment of candidate pathway variants.

At present, research reports on transcription factor-regulated biosensing systems have not been able to achieve targeted detection of high-value compounds and their analogs. Professor Huo Yixin 's team has addressed this problem and constructed a transcription factor BmoR-driven biosensing system for higher alcohols based on the transcription factor BmoR, which uses C2-C5 higher alcohols as signaling molecules. Due to the restricted characteristics of wild-type BmoR, it cannot achieve the specific detection of higher alcohol molecules, and the rational and irrational design and modification of the N-terminal functional region of BmoR are targeted.

The activation principle of wild-type BmoR is firstly analyzed, and the binding of wild-type BmoR to different alcohol molecules is simulated by molecular docking, which is used as a basis for fixed-point rational modification. By establishing a random mutant library and performing high-throughput screening of it, BmoR mutants specifically responding to n-butanol or isobutanol, respectively, are successfully obtained, which can maintain their specificity in the background noise of 0-800 mM (36.9 g/L) ethanol. In addition, a BmoR mutant with a highly sensitive response to higher alcohols, which is 107-fold more sensitive compared to the wild type, and a BmoR mutant with an extended detection range of 200 mM (14.8 g/L) are obtained. Through a series of design modifications, BmoR mutants with different properties are finally obtained, which can be used to construct a series of multifunctional biosensing systems for screening ideal strains or establishing dynamic control systems in the field of metabolic engineering.

Paper link:https://doi.org/10.1021/acssynbio.1c00549

With author's profile:

Professor Huo YiXin , long-appointed professor and doctoral supervisor in the School of Life Science, BIT. He has achieved a series of research results for industrial application by means of synthetic biology, mainly focusing on "microbial refining and manufacturing of bulk chemicals". His representative works have been published in top journals such as Science Nat Biotechnol, Nat Commun and so on. In the past five years, he has published more than 40 high-level papers and granted nearly ten international and domestic patents. He has presided over more than ten national, provincial and regional projects, such as the Natural Science Foundation of China and the National Key Research and Development Program. He is currently the director of the Chinese Society of Biological Engineering, the deputy director of the Key Laboratory of Molecular Medicine and Biological Diagnosis of the Ministry of Industry and Information Technology, the responsible professor of biotechnology of BIT .etc

Chen Zhenya , associate researcher in the School of Life Science, BIT. She has been engaged in the research of metabolic engineering and synthetic biology, biosensor mining and modification for a long period. She has published more than 20 papers in top journals such as Metabolic Engineering, ACS Synthetic Biology, Journal of Agricultural and Food Chemistry, Applied Microbiology and Biotechnology, Biotechnology and Bioengineering, and has been granted a number of invention patents.